The mission of Core A: Mass Spectrometry Core is to provide access to advanced mass spectrometry equipment for Pharmacology Center investigators and to provide assistance to investigators for development of novel analytical methods. The specific aims of this core are: Specific Aim 1 Develop ultra-high sensitivity LC-MS/MS protocols for characterization and quantitation of complex lipid and drug metabolite samples using nanoflow UPLC techniques and the proposed high sensitivity Quantum Vantage mass spectrometer. Specific Aim 2 - Develop sophisticated data-dependent scanning protocols for characterization of complex lipid and drug metabolite profiles for structural determination, similar to techniques employed by proteomics laboratories, using nanoflow LC and high resolution MS. Specific Aim 3 - Develop derivatization strategies to improve sensitivity for analysis of molecules with poor intrinsic ionization properties. These services will employ primarily liquid chromatography-tandem mass spectrometry (LC-MS/MS) for quantitation of biomolecules in tissues and fluids, identification of novel reactive oxidation products of lipids, and identification of protein adducts of reactive species. The core will use the MS equipment provided by the Vanderbilt University Mass Spectrometry Research Center, which includes a comprehensive small molecule mass spectrometry core (MS Core) and a state-of-the-art proteomics core. The instrumentation available to investigators includes a variety of low resolution (four triple quadrupole, two LTQ ion trap) and two high resolution (an Orbitrap FT-MS and a Q-TOF) LC-MS/MS instruments, many of which are equipped with ultra-high pressure liquid chromatographs for high efficiency separation of complex mixtures. In addition, we propose to purchase a new high sensitivity triple quadrupole MS equipped with a nanospray ion source and a nanoflow UPLC system. The combination of nanoflow LC with a state-of-the-art triple quadrupole MS will achieve about 100-fold greater sensitivity than currently possible with existing equipment. This increased performance will specifically benefit Project 1 (PI LJ Roberts), Project 2 (PI: JA Oates) and Project 5 (PI: AR Brash).